SPINE Protein Interaction Set

Features & Benefits

• Advantages:
• Only one tag (Strep-tag® II)
• Preys are covalently linked to the bait and weakly associated preys cannot escape detection
• High-stringency wash procedure reduces background
• Fast and easy one-step purification
• Time-resolved analysis of binding processes possible
• Convenient and highly flexible bait cloning procedure available (StarGate®)
• Disadvantages and Risks:
• At present validated for Strep-tag® II and prokaryotic expression systems only

Description

In this method the advantages of the One-STrEP purification system are combined with those of reversible in vivo protein cross-linking. Formaldehyde links proteins which are in close contact to each other thereby fixing the cellular protein:protein-interaction status at a given point of time. Stabilized protein complexes are purified on Strep-Tactin® and then resolved into components by short heating for further analysis.

SPINE (Strep-Protein Interaction Experiment12 with Strep-tag®II) has the possibility to covalently link preys to its bait by formaldehyde cross-linking. This linkage is achieved in the living organism enabling a time resolved snapshot of interacting proteins. SPINE is currently validated in prokaryotes only but its adaptation to mammalian systems is under way.